Accumulated experience towards recruiting a patient's immune system to battle malignant lymphomas suggest that polypeptide vaccines offer the best combination of safety and efficacy. The adenovirus approach proposed in the original project application for the treatment of T-cell lymphomas has been abandoned. However, we are now faced with the major challenge of producing these patient-specific polypeptide therapeutics rapidly and at an acceptable cost. A new and unique product is required for each patient. Recent and exciting advances in the Swartz laboratory in the Department of Chemical Engineering at Stanford suggest the feasibility of using cell-free protein synthesis technology. Preliminary results show reliable synthesis of a variety of vaccine candidates and also suggest promise for producing bioactive immune stimulators such as GMCSF. The research proposed in this application will continue to develop technology that will eventually lead to production of vaccines within a week of specimen acquisition and at costs that are a fraction of those required for competing technologies. These capabilities are essential for the general adoption of this promising therapy for treating T-cell malignancies. [unreadable] The specific aims for this project are: [unreadable] 1. Increase the stability of linear DNA templates in cell-free synthesis reactions to allow efficient vaccine production from PCR products. [unreadable] 2. Develop reliable and inexpensive technology for the consistent expression of 5 mg of insoluble vaccine polypeptide per batch reaction. [unreadable] 3. Develop reliable and inexpensive technology for the expression and purification of 5 mg of properly folded vaccine antigen per batch reaction. [unreadable] 4. Produce a variety of insoluble and soluble fusion proteins as vaccine candidates suitable for efficacy and safety tasting in animal models. [unreadable] This is a supplemental project to complement Project 2 of the Ronald Levy Program Grant entitled "Clinical and Laboratory Studies of Malignant Lymphomas" and focuses specifically on T-cell malignancies. It is required because of the abandonment of the adenovirus approach and is motivated by the opportunity presented by recent advances in cell-free protein synthesis technology. It is projected to have a two-year duration and will be directed by James R. Swartz, Professor, Department of Chem. Engineering, Stanford University. [unreadable] [unreadable] [unreadable]